Anti-CD40 [2C5]

Description:

Recombinant monoclonal antibody to CD40. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 2C5.

Clone Name:

2C5

Published Application(s):

agonist, immune activation, FC, IF

Published Species Reactivity:

Chicken

Products based on this clone:

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Ab01617-1.1 Anti-CD40 [2C5]
Chicken
Mouse IgG1
Purified
In Stock

Ab01617-23.0 Anti-CD40 [2C5]
Chicken
Rabbit IgG
Purified
In Stock

Alternative Name(s) of Target: CD40L receptor; CDw40; Bp50; B-cell surface antigen CD40; Tumor necrosis factor receptor superfamily member 5
Immunogen: This antibody was raised by immunising mice with a recombinant protein corresponding to the extracellular domain of chicken CD40 (chCD40ED).

Specificity: This antibody recognises extracellular domain of chicken CD40. CD40, an integral membrane glycoprotein of the TNF-receptor superfamily, is expressed mainly on professional antigen-presenting cells (APCs), including B-cells, macrophages, and dendritic cells. However, high levels of CD40 can also be detected on a wide range of tumor cells of the bladder, breast, and ovary.

Application Notes: 2C5 antibody is a CD40 agonist and can be used to induce immune activation. Agonistic monoclonal anti-CD40 antibodies (mAbs) mimic the effects of CD40L on APCs, leading to enhanced T-cell priming andexpansion, increased antibody production and class switching. This antibody was shown to be a powerful adjuvant eliciting a strong IgG-response in chicken what makes it an important agent in vaccines development (Chen et al., 2012). Furthermore, 2C5 was also demonstrated to be effective in detecting CD40 via immunofluorescence as well as via flow cytometry in chicken B cells and macrophages (Chen et al., 2010). Finally, this antibody was observed to activate NO synthesis in HD11 macrophages, and stimulate DT40 B-cellproliferation (Chent et al., 2010).

Antibody first published in: Chen et al. Production and characterization of agonistic monoclonal antibodies against chicken CD40. Dev Comp Immunol. 2010 Nov;34(11):1139-43. doi: 10.1016/j.dci.2010.06.014. Epub 2010 Jul 3. PMID:20599554
Note on publication: This article describes the generation and characterisation of 2C5 antibody.

IF staining of chicken peripheral blood monocytes using Ab01617-23.0. Click on image for more details.

Immunofluorescence staining of fixed chicken peripheral blood monocytes with anti-CD40 antibody 2C5 (Ab01617) Immunofluorescence analysis of paraformaldehyde fixed chicken peripheral blood monocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 2C5 (Ab01617-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01617-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.

Flow cytometry analysis of chicken peripheral blood monocytes using Ab01617-23.0. Click on image for more details.

Flow cytometry using the anti-CD40 antibody 2C5 (Ab01617). chicken peripheral blood monocytes were fixed using 2% PFA and stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG1 version of 2C5 (Ab01617-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

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